Control of Cellular Function by Reversible Photoregulation of Translation
Control of Cellular Function by Reversible Photoregulation of Translation
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DOI：
10.1002/cbic.201402495
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发表时间：
2014-12-15
期刊：
影响因子：
3.2
通讯作者：
Ogasawara, Shinzi
中科院分区：
文献类型：
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作者：
Ogasawara, Shinzi
使用光作为外部刺激提供了时空控制的潜力，因此是控制活细胞中基因表达的理想选择。在常用的笼蔽系统中，一旦笼蔽化合物被去除，由于解笼蔽反应的不可逆性，蛋白质表达就无法停止。我们开发了一种借助光响应帽来调节蛋白质表达的可逆方法，该帽可以通过触发帽的顺反光异构化以可逆的方式控制 mRNA 的翻译。在反式形式中，光响应帽完全抑制翻译，而顺式形式产生蛋白质（翻译的蛋白质是反式形式的 12.7 倍）。此外，我们成功地控制了活哺乳动物细胞中蛋白质表达的水平、时间和持续时间。此外，通过控制组成型活性 H-Ras 61L 蛋白表达来光诱导 PC12 细胞的神经元分化。
The use of light as an external stimulus offers the potential for spatiotemporal control and is thus ideal for controlling gene expression in living cells. In commonly used caging systems, once the caging compound is removed, protein expression cannot be stopped, due to the irreversibility of the uncaging reaction. We have developed a reversible method for regulating protein expression with the aid of a photoresponsive cap that can control the translation of mRNA in a reversible manner through triggering of cis-trans photoisomerization of the cap. In its trans form, the photoresponsive cap completely inhibited translation, whereas the cis form yielded protein (12.7 times more translated protein than in the trans form). Moreover, we succeeded in controlling the levels, timing and duration of protein expression in living mammalian cells. Additionally, neuronal differentiation of PC12 cells was photoinduced by controlling constitutively active H-Ras 61L protein expression.